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2
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1 #!/usr/bin/env python
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2
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50
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3 import argparse
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4 import logging
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5 from sys import stdout
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6 from subprocess import check_call
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7 from shutil import rmtree
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8 from tempfile import mkdtemp
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9 from os.path import isfile
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10 # avoid ugly python IOError when stdout output is piped into another program
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11 # and then truncated (such as piping to head)
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12 from signal import signal, SIGPIPE, SIG_DFL
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13 signal(SIGPIPE, SIG_DFL)
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14
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2
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15 tool_description = """
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16 Merge PCR duplicates according to random barcode library.
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17
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55
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18 Barcodes containing uncalled base 'N' are removed.
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2
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19
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20 Input:
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21 * bed6 file containing alignments with fastq read-id in name field
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8
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22 * fastq library of random barcodes
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2
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23
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24 Output:
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25 * bed6 file with random barcode in name field and number of PCR duplicates as
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26 score, sorted by fields chrom, start, stop, strand, name
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27
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28 Example usage:
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29 - read PCR duplicates from file duplicates.bed and write merged results to file
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30 merged.bed:
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31 merge_pcr_duplicates.py duplicates.bed bclibrary.fa --outfile merged.bed
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2
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32 """
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33
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34 epilog = """
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35 Author: Daniel Maticzka
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36 Copyright: 2015
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37 License: Apache
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38 Email: maticzkd@informatik.uni-freiburg.de
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39 Status: Testing
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40 """
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41
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42 # parse command line arguments
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43 parser = argparse.ArgumentParser(description=tool_description,
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44 epilog=epilog,
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45 formatter_class=argparse.RawDescriptionHelpFormatter)
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46 # positional arguments
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47 parser.add_argument(
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48 "alignments",
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49 help="Path to bed6 file containing alignments.")
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50 parser.add_argument(
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51 "bclib",
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52 help="Path to fastq barcode library.")
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2
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53 # optional arguments
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54 parser.add_argument(
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55 "-o", "--outfile",
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55
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56 required=True,
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2
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57 help="Write results to this file.")
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58 # misc arguments
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59 parser.add_argument(
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60 "-v", "--verbose",
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61 help="Be verbose.",
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62 action="store_true")
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63 parser.add_argument(
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64 "-d", "--debug",
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65 help="Print lots of debugging information",
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66 action="store_true")
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67 parser.add_argument(
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68 '--version',
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69 action='version',
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70 version='0.2.0')
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2
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71
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72 args = parser.parse_args()
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73
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74 if args.debug:
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75 logging.basicConfig(level=logging.DEBUG, format="%(asctime)s - %(filename)s - %(levelname)s - %(message)s")
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76 elif args.verbose:
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77 logging.basicConfig(level=logging.INFO, format="%(filename)s - %(levelname)s - %(message)s")
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78 else:
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79 logging.basicConfig(format="%(filename)s - %(levelname)s - %(message)s")
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80 logging.info("Parsed arguments:")
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81 logging.info(" alignments: '{}'".format(args.alignments))
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82 logging.info(" bclib: '{}'".format(args.bclib))
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83 if args.outfile:
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84 logging.info(" outfile: enabled writing to file")
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85 logging.info(" outfile: '{}'".format(args.outfile))
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86 logging.info("")
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87
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88 # see if alignments are empty and the tool can quit
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89 n_alns = sum(1 for line in open(args.alignments))
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90 if n_alns == 0:
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91 logging.warning("WARNING: Working on empty set of alignments, writing empty output.")
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92 eventalnout = (open(args.outfile, "w") if args.outfile is not None else stdout)
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93 eventalnout.close()
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94 exit(0)
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95
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96 # check input filenames
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97 if not isfile(args.bclib):
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98 raise Exception("ERROR: barcode library '{}' not found.")
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99 if not isfile(args.alignments):
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100 raise Exception("ERROR: alignments '{}' not found.")
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101
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102 try:
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103 tmpdir = mkdtemp()
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104 logging.debug("tmpdir: " + tmpdir)
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2
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105
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106 # prepare alinments
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107 syscall2 = "cat " + args.alignments + " | awk -F \"\\t\" 'BEGIN{OFS=\"\\t\"}{split($4, a, \" \"); $4 = a[1]; print}'| sort --compress-program=gzip -k4,4 > " + tmpdir + "/alns.csv"
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108 check_call(syscall2, shell=True)
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2
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109
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110 # join barcode library and alignments
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111 syscall3 = "cat " + args.bclib + " | awk 'BEGIN{OFS=\"\\t\"}NR%4==1{gsub(/^@/,\"\"); id=$1}NR%4==2{bc=$1}NR%4==3{print id,bc}' | sort --compress-program=gzip -k1,1 | join -1 1 -2 4 - " + tmpdir + "/alns.csv " + " | awk 'BEGIN{OFS=\"\\t\"}$4!~/N/{print $3,$4,$5,$2,$6,$7}' | datamash --sort -g 1,2,3,4,6 count 4 | awk 'BEGIN{OFS=\"\\t\"}$4!~/N/{print $1,$2,$3,$4,$6,$5}' > " + args.outfile
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112 # 'chrom', 'start', 'stop', 'bc', 'ndupes', 'strand'
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113 check_call(syscall3, shell=True)
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114 finally:
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115 logging.debug("removed tmpdir: " + tmpdir)
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116 rmtree(tmpdir)
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